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Preparation of DNA for Pulsed-field Gel Electrophoresis: Isolation of DNA from Mammalian Cells - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4030

Category: Nucleic Acid Electrophoresis Protocols: Pulsed Field Gel Electrophoresis Protocols

Protocol for preparation of DNA for pulsed-field gel electrophoresis: isolation of DNA from mammalian cells and tissues. Genomic DNAs from mammalian cells are prepared for pulsed-field gel electrophoresis by lysing cells in situ in an agarose plug. Following digestion with an appropriate restriction enzyme, the plug is loaded directly into the well of a pulsed-field gel or it can be melted before loading. - [Read Preparation of DNA for Pulsed-field Gel Electrophoresis: Isolation of DNA from Mammalian Cells]

Preparation of DNA for Pulsed-field Gel Electrophoresis: Isolation of Intact DNA from Yeast - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3235

Category: Nucleic Acid Electrophoresis Protocols: Pulsed Field Gel Electrophoresis Protocols

Protocol for preparation of DNA for pulsed-field gel electrophoresis: isolation of intact DNA from yeast. Yeast cells are first treated enzymatically to break down the cell walls and then resuspended in low-melting-temperature agarose plugs. The DNA is liberated by infusing the plugs with lysis buffer and proteases. This method is used to prepare both conventional and artificial yeast chromosomes. - [Read Preparation of DNA for Pulsed-field Gel Electrophoresis: Isolation of Intact DNA from Yeast]

Preparation of DNA from Cells in 24-Well Tissue Culture Plates Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/18/pdb.prot4414

Category: Stem Cell Protocols

This protocol describes a method of DNA preparation from ES cells in 24-well plates. - [Read Preparation of DNA from Cells in 24-Well Tissue Culture Plates Protocol]

Preparation of Double-stranded (Replicative Form) Bacteriophage M13 DNA Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3281

Category: Viral Manipulation Protocols: M13 Phage Manipulation Protocols

The double-stranded replicative form (RF) of bacteriophage M13 is isolated from infected cells using methods similar to those used to purify plasmid DNA. Several micrograms of RF DNA can be isolated from a 1-2-ml culture of infected cells. - [Read Preparation of Double-stranded (Replicative Form) Bacteriophage M13 DNA Protocol]

Preparation of Embryonic Stem (ES) Cells for Injection Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/21/pdb.prot4416

Category: Stem Cell Protocols: Stem Cell Injection Protocols

Injection of ES cells into the cavity of blastocysts is one of the main methods for generating chimeras. This protocol describes the preparation of ES cells for injection. - [Read Preparation of Embryonic Stem (ES) Cells for Injection Protocol]

Preparation of Endothelial Cells Protocol - https://catalog.invitrogen.com/index.cfm?fuseaction=iProtocol.unitSectionTree&treeNodeID=E538DDD4A302A0DF2CDDD9363072C1CA&objectid=6673B460F50B90057960CE47ABB708A4

Category: Cell Biology and Cell Culture: Endothelial Cell Protocols

Endothelial cells, which line blood vessels, can be prepared from a variety of tissues. They are frequently prepared from the umbilical vein, which is relatively easy to obtain. The procedure is clearly described and provides a large population of highly purified endothelial cells. There are also methods for obtaining endothelial cells from other tissues such as fat, skin, and mucosa. These methods require special care and generate smaller populations of cells. - [Read Preparation of Endothelial Cells Protocol]

Preparation of Frozen Stocks of RAW 264.7 Cells - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000180.pdf?pid=PP00000180

Category: Cell Culture Protocols: Cell Lines Protocols

Protocol for the preparation of frozen stocks of RAW 264.7 cells. Includes: Freezing procedure and reagents. - [Read Preparation of Frozen Stocks of RAW 264.7 Cells]

Preparation of Genomic DNA from Mouse Tails and Other Small Samples Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4038

Category: Nucleic Acid Purification Protocols: DNA Isolation Protocols: Genomic DNA Isolation Protocols

Simple protocol is used to extract DNA from small numbers of cultured cells and from fragments of soft or bony tissues. The method is used chiefly to genotype transgenic and knockout mice. Each 6-10-mm snippet of mouse tail yields 50-100 µg of DNA that can be used in dot or slot blotting to detect a transgene of interest, in Southern hybridization to detect DNA fragments that are <20 kb in size, and as a template in PCRs. - [Read Preparation of Genomic DNA from Mouse Tails and Other Small Samples Protocol]

Preparation of Genomic DNA from Yeast Using Glass Beads Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4151

Category: Yeast Protocols: Yeast Nucleic Acid Protocols: Yeast DNA Isolation Protocols

Protocol describes the preparation of genomic DNA from a 5-ml liquid culture of yeast cells. - [Read Preparation of Genomic DNA from Yeast Using Glass Beads Protocol]

Preparation of High Titer Adenovirus in P11 Cells Protocol - http://www.whitelabs.org/Lab%20Protocols/adenovirus%20protocols/Preparation%20of%20High%20Titer%20Adenovirus%20in%20P11%20cells.htm

Category: Viral Culture and Isolation Protocols: Viral Isolation Protocols

Protocol for the preparation of high titer adenovirus in P11 cells. - [Read Preparation of High Titer Adenovirus in P11 Cells Protocol]

Preparation of KC Nuclear Extract for In Vitro Splicing - http://www.bio.com/protocolstools/protocol.jhtml%3Bjsessionid%3D34E3CBZRBQMFZR3FQLMCFEWHUWBNSIV0?id=p521

Category: Cell Biology and Cell Culture: In Vitro Reconstitution Protocols

Protocol for preparation of KC nuclear extract for in vitro splicing. Protocol makes 3.4 ml of extract for every 4 liter of cells (depending on initial cell concentration). Protocol includes: Procedure, Solutions, BioReagents and Chemicals and protocol hints. - [Read Preparation of KC Nuclear Extract for In Vitro Splicing]

Preparation of Mouse Embryonic Fibroblast (MEF) Feeder Plates Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/18/pdb.prot4483

Category: Stem Cell Protocols: Feeder Cell Preparation Protocols

MEF feeders are prepared weekly to provide a substrate for undifferentiated embryonic stem (ES) cells. Primary MEF cells are thawed, established in culture, treated with mitomycin C to halt their proliferation so they cannot overgrow the ES cultures, and then replated onto dishes convenient for ES cell culture. This protocol can also be used to prepare feeder cells from STO fibroblast cell lines. - [Read Preparation of Mouse Embryonic Fibroblast (MEF) Feeder Plates Protocol]

Preparation of Myocyte Lysates for Cyclic AMP Determination - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000131.pdf?pid=PP00000131

Category: Signalling Signal Transduction Protocols

Protocol provides a method for acheiving a sufficient sample for several determinations of cAMP. The protocol described for measuring the content of cyclic adenosine 3',5'- monophosphate (cyclic AMP or cAMP) in cardiac myocytes is an enzyme-linked immunoassay system. Protocol includes information on: Treatment of Cells and Preparation of Extracts; Use of Environmental Chamber; Reagents and Materials. - [Read Preparation of Myocyte Lysates for Cyclic AMP Determination]

Preparation of Nuclear Extracts for Gel Shifts and Westerns Blots - http://faculty.virginia.edu/mirmira/resources_files/Protocols/Nuclear%20Extracts.htm

Category: Protein Protocols: Nuclear Extraction Protocols

Nuclear Extract protocol is optimized for about 10^7 cells, can use for gel shift assays and western blotting. Mirmira Lab, Univ. Virginia - [Read Preparation of Nuclear Extracts for Gel Shifts and Westerns Blots]

Preparation of RAW 264.7 Lysates for Cyclic AMP Determination - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000175.pdf?pid=PP00000175

Category: Signalling Signal Transduction Protocols

The protocol provides a method to achieve a sample sufficient for one determination of cAMP using the acetylation protocol. The protocol describes the method used for measuring the content of cyclic adenosine 3',5'- monophosphate (cyclic AMP or cAMP) in RAW 264.7 cells using an enzyme-linked immunoassay system. Information included in the protocol: Treatment of Cells and Preparation of Extracts; Reagents and Materials. - [Read Preparation of RAW 264.7 Lysates for Cyclic AMP Determination]

Preparation of RNA from Plant Tissue Using Trizol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4105

Category: Plant Biology Protocols: Plant DNA RNA Isolation Protocols

Protocol is based on the standard Trizol protocol for the purification of RNA from animal cells using Trizol (Purification of RNA from Animal Cells using Trizol). In this version, adapted for use with plant tissues, a high-salt isopropanol precipitation step has been added to precipitate RNA selectively, while maintaining polysaccharides and proteoglycans in solution. - [Read Preparation of RNA from Plant Tissue Using Trizol]

Preparation of Single-stranded Bacteriophage M13 DNA Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3686

Category: Viral Manipulation Protocols: M13 Phage Manipulation Protocols

Bacteriophage M13 single-stranded DNA is prepared from virus particles secreted by infected cells into the surrounding medium. The filamentous particles are concentrated by precipitation from a high-ionic-strength buffer with polyethylene glycol. Subsequent extraction with phenol releases the single-stranded DNA, which is then collected by precipitation with ethanol. This protocol is generally used to prepare single-stranded DNA from a small number of M13 isolates. - [Read Preparation of Single-stranded Bacteriophage M13 DNA Protocol]

Preparation of ultra-competent E. coli cells for transformation - http://www.hybtech.org/Liu/uccell.html

Category: Microbiology: Bacterial Competent Cells Preparation E.Coli

Preparation of ultra-competent E. coli cells for transformation. Dr. Chun-Ming Liu. Based on Inoue et al (1990). - [Read Preparation of ultra-competent E. coli cells for transformation]

Preparation of whole cell homogenates of cultured mammalian cells for 2-D Electrophoresis - http://www.healthsystem.virginia.edu/internet/biomolec/2dgelsampleprep.cfm

Category: Proteomic Protocols

Preparation of whole cell homogenates of cultured mammalian cells for 2-D Electrophoresis. Univ. Virginia. - [Read Preparation of whole cell homogenates of cultured mammalian cells for 2-D Electrophoresis]

Preparation of Yeast Cells for Flow Cytometry Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4178

Category: Yeast Protocols: Yeast Nucleic Acid Protocols

Flow cytometry is used to analyze the quantity of DNA in cells. Since the DNA content of cells varies through the cell cycle, this information can provide an indication of cell cycle progression. This protocol uses SYTOX Green staining. - [Read Preparation of Yeast Cells for Flow Cytometry Protocol]

Preparing Agrobacterium Cells That Simplifies the Arabidopsis Transformation Protocol - http://www.plantmethods.com/content/2/1/16

Category: Plant Biology Protocols: Arabidopsis Transformation Protocols

Protocol for an improved method for preparing Agrobacterium cells that simplifies the Arabidopsis transformation protocol. - [Read Preparing Agrobacterium Cells That Simplifies the Arabidopsis Transformation Protocol]

Preparing Cells for PI/FACS (cell cycle) Analysis Protocol - http://www.flemingtonlab.com/Protocols/PreparingCellsforFACS-PI.pdf

Category: Cell Biology and Cell Culture: Cell Cycle Protocols

This method works well to assess cell cycle distribution of whole cell populations. This method can also be used to assess the cell cycle distribution of GFP transfected cells however, the EtOH step is generally not sufficient to keep GFP in the cell. - [Read Preparing Cells for PI/FACS (cell cycle) Analysis Protocol]

Preparing Cells for PI/FACS (cell cycle) Analysis Protocol - http://www.flemingtonlab.com/Protocols/PreparingCellsforFACS-PI.pdf

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Cell Cycle Analysis Protocols

Protocol for preparing cells for PI/FACS (cell cycle) analysis. - [Read Preparing Cells for PI/FACS (cell cycle) Analysis Protocol]

Preparing Cells for PI/FACS (cell cycle) Analysis Protocol - http://www.flemingtonlab.com/Protocols/PreparingCellsforFACS-PI.pdf

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Flow Cytometry Data Analysis Protocols

Protocol for preparing cells for PI/FACS (cell cycle) analysis. - [Read Preparing Cells for PI/FACS (cell cycle) Analysis Protocol]

Preparing Chemically Competent Cells - http://openwetware.org/wiki/Preparing_chemically_competent_cells

Category: Microbiology: Bacterial Competent Cells Preparation E.Coli

Detailed protocol Preparation and Steps for E.Coli Competent Cells procedure. Open Net Ware. - [Read Preparing Chemically Competent Cells]

Articles (1-10 of 24)

Lambda Phage Protocol Large DNA Preparation

Single Stranded Plasmid DNA Isolation Protocol

A Single Stranded Plasmid DNA Isolation Protocol describing the production and isolation of single-stranded DNA (ssDNA) using bacteriophagemid-containing bacteria and helper phage. Infection of the host cells with helper phage allows for packaging of ssDNA into bacteriophage. The ssDNA can then be isolated from phage particles.

Genomic DNA Labeling of Microarrays Protocol

DNA microarrays are an ordered arrangement of DNA molecules complementary to genes of interest that are "spotted" by robotic equipment onto a glass slide substrate. The expression of genes in cells can be monitored with microarrays by preparing cDNA from the mRNA of cells of interest and measuring the hybridization to the microarray. This protocol describes the labeling of genomic DNA for use as a probe for hybridization to the cDNA spotted on the array.

Drosophila Cell Transfection Protocol

A simple Drosophila tissue culture cell transfection method.

Tubulin Polymerization Protocol using GTP and GMPCPP

Tubulin is polymerized into microtubules by incubating tubulin at 37°C with GTP. A nucleation seed is added when the purpose is to assay microtubule elongation. Tubulin can also be polymerized for the purposes of recycling the tubulin or labeling the microtubules with fluorescently labeled tubulin. Based on the protocol by Timothy Mitchison of Harvard University.

Microarray Protocol Preparation of Fluorescent DNA Probes from Human mRNA

This Microarray Protocol Preparation of Fluorescent DNA Probes from Human mRNA protocol describes the production of probes labeled with the fluorescent dyes, Cy3 and Cy5, following the synthesis of cDNA from human mRNA and the hybridization of the probes to DNA microarrays.

Acid Guanidinium Thiocyanate RNA Isolation Phenol Chloroform Extraction

A single step RNA isolation protocol using Phenol Chloroform Extraction and Acid Guanidinium Thiocyanate. This RNA isolation method uses the fact that guanidinium thiocyanate can simultaneously lyse the cells and inactive cellular RNAses during the initial RNA isolation step allow a single step in the method.

Microinjection Pipettes Protocol

Microinjection Pipettes Preparation Protocol.

Acid Washing Microinjection Pipettes Protocol

Acid Washing of Glass microinjection pipettes protocol.

Paraffin Embedding Protocol

Paraffin Embedding Protocol for molecular profiling. This Paraffin Embedding Protocol describes the processing of the tissues into sections following ethanol fixation. Molecular profiling (MP) is a technique that is used to visualize the global patterns of RNA expression or protein expression in various cell types and disease processes.

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